Tuft cell heterogeneity in function, lineage, and structure in ileal inflammatory disease

项目概要/摘要 炎症性肠病（IBD）及其亚型克罗恩病（CD）是由于缺乏免疫抑制剂而引起的。 遗传易感个体对环境抗原的耐受性纵向分析 已经确定了传染性寄生虫病和IBD之间的负相关性， 这一现象被称为“卫生假说”簇细胞最近被鉴定为 是感应和应对寄生虫感染的关键部件。我们的实验室最近 发现簇状细胞在其谱系和微生物组中的功能上可能是异质的， 反应，并且特定人群对于减轻回肠炎症中的炎症可能至关重要 疾病在这里，我们提出了创新的实验和计算方法来剖析 与调节免疫相关簇细胞谱系、结构和功能的异质性 在CD的体内模型中的反应。在目标1中，我们将进行单细胞测序， 分辨率显微镜和功能性分泌测定，以了解谱系结构功能 小鼠和肠模型中不同簇状细胞群之间的关系。在目标2中， 多重成像、流式细胞术飞行时间（CyTOF）和细胞因子分析，以探测 下游簇2型响应网络，利用几个化学和遗传模型， 通路消融通过我们的研究结果，我们的目标是为当前的 了解肠道中的簇细胞生物学和上皮-微生物组串扰。最后， 这项研究将使我们能够诊断、分层并最终治疗IBD患者， 这种复杂、多因素疾病的靶点库。

PROJECT SUMMARY/ABSTRACT Inflammatory Bowel Disease (IBD) and its subtype Crohn’s disease (CD) arise due to a loss of tolerance to environmental antigens in genetically susceptible individuals. Longitudinal analysis has identified an inverse correlation between rates of communicable parasitic diseases and IBD, a phenomenon termed the “hygiene hypothesis.” Tuft cells have recently been identified as a critical component for sensing and responding to parasitic infections. Our lab has recently identified that tuft cells may be heterogeneous in their lineage and function in microbiome response, and specific populations may be critical for mitigating inflammation in ileal inflammatory disease. Here, we propose innovative experimental and computational approaches to dissect the heterogeneity of tuft cell lineage, structure, and function as they relate to regulating immune responses in in vivo models of CD. In aim 1, we will conduct single-cell sequencing, high resolution microscopy, and functional secretory assays to understand lineage-structure-function relationships in different tuft cell populations in mouse and enteroid models. In aim 2, we utilize multiplex imaging, Cytometry Time-of-Flight (CyTOF), and cytokine profiling to probe the downstream tuft-type 2 response network by utilizing several chemical and genetic models of pathway ablation. Through our findings, we aim to make significant contributions to the current understanding of tuft cell biology and epithelial-microbiome crosstalk in the intestine. Ultimately, this research will allow us to diagnose, stratify, and, ultimately, treat patients with IBD and expand the pool of targets in this complex, multifactorial disease.